HIV Infections ; diagnosis ; therapy ; Humans ; Medicine, Chinese Traditional ; methods

Objective: To explore the characteristics of IR spectra of the crude drug Herba Sedi collected in different habitats and seasons.Methods: The infrared spectra of all the samples of the whole plants were obtained by FT-IR spectroscopy,the sample powder tablets were used,and identifi cation features were determined through comparative study.Results: The harvest seasons of the samples are remarkly related to the positions(cm-1),intensity or ratios of some major absorbing peaks of the infrared spectra from the 15 samples of several different months and 4 habitats in Hubei Province,China.As for peak positions,the peak values(1625?7)cm-1 of all the samples in spring are less than 1623cm-1,but in autumn(April to October) are generally more than 1628 cm-1.And peak values(1057?16)cm-1 of the samples of spring are more than 1050 cm-1,but less than 1050 cm-1 for the samples of summer and autumn.Moreover,the habitats of samples are also related to IR spectra to some certain extent.Conclusion: The harvest seasons of Herba Sedi from Hubei Province can be preliminarily ascertained according to the IR spectra features.IR technique should be paid attention to in research and identifi cation of the harvest periods of crude plant drugs.

Objective To investigate the effects of radiation on the apoptosis rate and expression of 7 apoptosis-related genes in well-differentiated human nasopharyngeal carcinoma cell line CNE-1.Methods CNE-1 cells were cultured in vitro.The apoptosis rates of CNE-1 cells under 0-,2-,4-,6-,and 8-Gy radiation were measured by flow cytometry.The mRNA expression levels of Bcl-2,Bcl-xl,Bcl-w,Bax,Bak,Bad,and Bid were measured by RT-PCR.The Pearson test was used for analyzing the correlation of mRNA expression with apoptosis rate and radiation dose and the correlation between apoptosis rate and survival fraction.Results The early apoptosis rate of CNE-1 cells increased gradually as the radiation dose ranged from 0 to 6 Gy,but decreased when the radiation dose was 8 Gy; the late apoptosis rate of CNE-1 cells increased as the radiation dose ranged from 0 to 8 Gy.The mRNA expression of Bax was upregulated as CNE-1 cells were irradiated,and it was positively correlated with the early/late apoptosis rate and radiation dose (P =0.000 for all comparisons).The mRNA expression of Bcl-xl was downregulated,and it was negatively correlated with the early/late apoptosis rate (P =0.005 and 0.039) ; but it showed no correlation with radiation dose (P =0.369).The mRNA expression of Bcl-2 was upregulated and reached the peak level when the radiation dose was 4 Gy,and then it fell as the radiation dose increased.The mRNA expression of Bcl-w,Bcl-2,Bad,and Bid were not correlated with the early/late apoptosis rate (P =0.058 -0.894).There was no correlation between the apoptosis rate and survival fraction in CNE-1 cells (P =0.064).Conclusions Bax and Bcl-xl have some correlation with apoptosis rate in CNE-1 cells,but no correlation between the apoptosis rate and survival fraction was observed.

Objective:To investigate the correlation between white matter hyperintensities (WMHs) and stroke etiology classification in patients with acute isolated penetrating artery territory infarction.Methods:Patients with first-ever acute isolated penetrating artery territory infarction admitted to the Department of Neurology, the Affiliated Hospital of Xuzhou Medical University from January 2017 to May 2020 were enrolled retrospectively. According to the Chinese Ischemic Stroke Subclassification (CISS) system, they were divided into large artery atherosclerosis (LAA) and perforating artery disease (PAD). According to the distribution of infarcts, they were divided into lenticulostriate artery (LSA) territory infarction and paramedian pontine artery (PPA) territory infarction. The demographics, vascular risk factors, baseline clinical data, WMHs location, and Fazekas Scale scores were documented. Multivariate logistic regression analysis was used to identify the independent influencing factors of stroke etiology classification. Results:A total of 440 patients with acute isolated penetrating artery territory infarction were enrolled, including 120 (27.3%) in the LAA group, and 320 (72.7%) in the PAD group; 213 (48.4%) with LSA territory infarction, and 227 (51.6%) with PPA territory infarction. The proportion of patients with total Fazekas score 3-6 and periventricular WMHs (PWMHs) score 2-3 in the PAD group was significantly higher than those in the LAA group (all P<0.05). In patients with LSA territory infarction, the proportion of the patients with hypertension, WMHs total Fazekas score 3-6 and PWMHs score 2-3 in PAD subgroup was significantly higher than those in the LAA subgroup, while the proportion of the patients with hyperlipidemia was significantly lower than that in LAA subgroup (all P<0.05). In patients with PPA territory infarction, the levels of low-density lipoprotein cholesterol and homocysteine in the PAD subgroup were significantly lower than those in the LAA subgroup. Multivariate logistic regression analysis showed that PWMHs score 2-3 was an independent correlation factor of PAD (odds ratio [ OR] 2.220, 95% confidence interval [ CI] 1.085-4.541; P=0.029). In patients with LSA territory infarction, hyperlipidemia was independently correlated with LAA ( OR 0.432, 95% CI 0.192-0.972; P=0.042), and PWMHs score 2-3 was independently correlated with PAD ( OR 3.846, 95% CI 1.193-12.397; P=0.024). In patients with PPA territory infarction, higher low-density lipoprotein cholesterol ( OR 0.660, 95% CI 0.494-0.883; P=0.005), homocysteine ( OR 0.958, 95% CI 0.930-0.987; P=0.005) and C-reactive protein ( OR 0.987, 95% CI 0.977-0.997; P=0.008) were independently correlated with LAA. Conclusions:WMHs are common in patients with acute isolated perforating territory infarction caused by LAA and PAD, and more severe PWMHs suggest that PAD is more likely to be the cause of the acute isolated perforating territory infarction, especially in patients with LSA territory infarction.

The internal medicine of paediatrics is the important major course in paediatrics. Focusing on strengthening the constructionof staff, we cultivate all levels of teachers from different administrative level and personality, do schorlarly research meticulously,standardizing management and improve the teaching evaluation system. Besides we promote the construction and development of theinternal medicine of paediatrics by carrying out teaching research, impsoving teaching methods and making paediatics the key subjectin China.

Objective To investigated the role of antithrombin Ⅲ (AT-Ⅲ) levels in the early diagnosis of disseminated intravascular coagulation (DIC) in patients with sepsis and the predictive effect of AT-Ⅲ on the development of DIC.Methods A retrospective study was conducted. Patients admitted to intensive care unit (ICU) of the First Affiliated Hospital of China Medical University from January to December in 2015 were enrolled. The patients were divided into sepsis group and non-sepsis group according to the diagnostic criteria of sepsis. In addition, sepsis patients were divided into 3 subgroups according to the international society on thrombosis and haemostasis (ISTH) scores on the first day: overt DIC (ISTH ≥ 5), non-overt DIC (ISTH 1-4) and none DIC group (ISTH = 0). Blood routine test, prothrombin time (PT), fibrinogen (Fib), D-dimer, fibrin degradation products (FDP), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores, sequential organ failure assessment (SOFA) scores and ISTH scores were recorded on the first ICU day. AT-Ⅲ was recorded during 7 days. The differences were compared among these 3 groups. Correlations of AT-Ⅲ with various parameters were calculated by using Pearson correlation analysis in sepsis group and overt DIC group. Receiver operating characteristic (ROC) curves for diagnosis of DIC with AT-Ⅲ, AT-Ⅲ+PT were drawn to evaluate the diagnostic efficiency. The AT-Ⅲ levels of DIC patients were compared between early-onset DIC and late-onset DIC during their ICU stay. The change of AT-Ⅲ levels with time and prognosis in patients with early-onset DIC was compared between groups.Results Totally 445 patients were recruited, with 138 patients in sepsis group, and 307 in non-sepsis group. There were 20 patents diagnosed with overt DIC on the first ICU day, 115 patients non-overt DIC and 3 patients of none DIC. Twenty-five sepsis patients were diagnosed overt DIC during the ICU days. AT-Ⅲ level in sepsis patients on the first ICU day were lower than that in non-sepsis patients [(55.29±13.92)% vs. (76.54±12.31)%,P < 0.01]. Patients with overt DIC had a lower AT-Ⅲ level than non-overt DIC or none DIC patients [(43.85±13.00)% vs. (56.95±13.03)%, (68.00±16.52)%, bothP < 0.05]. It was shown by Pearson correlation analysis that AT-Ⅲ level of sepsis patients on the first ICU day was negatively correlated to ISTH score and PT (r value were -0.467, -0.654, bothP < 0.01). AT-Ⅲ level of overt DIC patient on the first ICU day was negatively correlated with PT (r = -0.675,P = 0.001). It was shown by ROC curve that area under ROC curve (AUC) of AT-Ⅲ combined with PT for diagnosis overt DIC in sepsis patients was higher than that of AT-Ⅲ or PT alone (0.843 vs. 0.763, 0.834), the sensitivity 90.0%, specificity 73.7%. The cut-off value for overt DIC diagnosis in sepsis patients of AT-Ⅲ level alone was 48.5%, sensitivity was 78.0%, specificity was 70.0%. On the first ICU day, AT-Ⅲ level was risen when ISTH score improved in patients with sepsis. There was similar change of AT-Ⅲ level between patients with early-onset DIC and late-onset DIC. AT-Ⅲ level increased with DIC improvement.Conclusion AT-Ⅲ level can be used for diagnosing sepsis-associated overt DIC independently or with a combination of PT. When ISTH score improved, AT-Ⅲ level was risen in patients with sepsis associated DIC.

Objective To investigate the expression of recombinant IL-37b protein and removal of the endotoxin, and identify its biological activity.Methods The prokaryotic expression vector pET28/IL-37b was constructed and to transform Escherichia coli (E.coli) Rosetta.After induction with IPTG, the recombinant protein was purified through Ni2+-NTA gel column and identified by SDS-PAGE and Coomassie brilliant blue staining.Then, the endotoxin protein was removed and was treated with LPS-stimulated RAW 264.7 cells.The culture supernatant was collected.The expression of IL-6 was detected by ELISA and the biological activity of the protein was identified.Results The recombinant IL-37b with high purity was expressed and the endotoxin produced by prokaryotic expression was reduced, and it was identified to have good biological activity.Conclusions In this study a recombinant IL-37b protein with high biological activity is successfully obtained.

Objective To evaluate efficacy of routine cleaning and disinfection methods for incubators,and put forward a feasible improvement solution.Methods 30 incubators used in a neonatal intensive care unit of a hospital between Decem-ber 2013 and June 2014 were chosen and randomly divided into baseline,control,and trial groups(10 incubators in each group).Baseline group and control group were disinfected by routing disinfection method (wiping internal and external sur-faces of incubators with water and chlorine-containing disinfectant),trial group adopted intensified disinfection method (wi-ping internal surfaces of incubators with alcohol)on the basis of routine disinfection,disinfectant efficacy of three groups were compared.Results In baseline group,unqualified incubators were initially detected on the fourth day of monitoring, all incubators were contaminated in varying degrees on the seventh day of monitoring,the detection rate of unqualified spec-imens was 31.43% (88/280).The median time for the initial detection of unqualified incubators in control group and trial group were on the fifth day and seventh day respectively,there was significant difference between two groups(χ2 =12.38, P <0.05);The unqualified rate of trial group was significantly lower than control group (15.36%[43/280]vs 32.86%[92/280],χ2 =23.43,P <0.05 ).Conclusion Intensified disinfection with alcohol on the basis of routine disinfection method can effectively improve the disinfectant efficacy of the surface of incubators,it is convenient,inexpensive and safe, and worth to be popularized in primary hospitals.

BACKGROUND: The study of isolation, purification, culture, cell labeling, inducing factors, effects of gene transfection on cytobiology, cell carrier construction, and time window for back transplantation of cell compound pertaining to marrow stromal cells (MSCs) is still in its infancy. OBJECTIVE: To search for an in vitro culture method that can be simply and effectively obtained. DESIGN, TIME AND SETTING: The present cytological in vitro experiment was performed at the Beijing Institute of Genome, Chinese Academy of Sciences between June 2006 and July 2007. MATERIALS: Eight specific pathogen-free New Zealand rabbits, aged 6 weeks, were provided by the Laboratory Animal Center, Institute of Genetics and Development, Chinese Academy of Sciences. METHODS: Under sterile condition, 1 mL rabbit bone marrow was taken and diluted with D-Hanks solution. Following centrifugation and subsequent supernatant removal, bone marrow was re-suspended using dulbecco's modified eagle's medium (DMEM) for single cell suspension. Next, single cell suspension was dropped onto the liquid surface of equal-volume lymphocyte separation medium (density: 1.077). Subsequent to centrifugation, cloudlike mononuclear cell layer was collected and re-suspended with DMEM containing 20% fetal bovine serum. The cells were inoculated at lxl0/cm2 and purified by adherent method. When 70%-80% of flask bottom was covered, cell digestion and passage was performed.MAIN OUTCOME MEASURES: Cell growth was observed with an operating microscope. Surviving cells were counted by Trypan blue viability test. Cell identification was performed by hematoxylin-eosin staining. Through the use of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, cell viability was detected to observe the cellular resuscitation of the cultured cells following cryopreservation. RESULTS: Twenty-four hours after inoculation, cells began to adhere to the wall, exhibiting short shuttle- or triangle-shaped appearance with different sizes of cellular processes. Three days later, adherent cells began to divide, and cell clusters could be found in some areas; One week later, most of cells exhibited scattered fibroblast-like growth; After passage, cells were evenly distributed with long shuttle-shaped appearance and arranged orderly. Following 3 passages, there wound be 5×106-1×107 adherent cells in 1 mL MSC suspension. Approximately 90% of MSCs survived and identified as mononuclear cells. Cells vigorously grew at days 1-6 after inoculation and reached a peak level at day 8, followed by a viability decline. After 56 days of resuscitation, frozen cells displayed a good and rapid growth. CONCLUSION: Highly purified MSCs can be acquired by gradient centrifugation of lymphocyte separation medium. Enough seeded cells can be obtained by in vitro culture and the cellular viability does not change after frozen preservation and resuscitation.

BACKGROUND: SOX-9 plays an important role in occurrence and development of cartilaginous tissues,enhances agglutination of mesenchymal cells,has structural domains of transcriptional activation,and can directly regulate the transcription,OBJECTIVE: To construct pDC316-SOX-9 plasmid for transfection of rabbit bone marrow stromal cells (BMSCs) using SOX-9 gene,and to investigate the effects of SOX-9 gene on growth characteristics of BMSCs and product expression. DESIGN,TIME AND SETTING: The cell gene engineering in vitro experiment was performe,d at the Beijing Institute of Genome, Chinese Academy of Scienees from June 2006 to January 2007.MATERIALS: Eight SPF New Zealand rabbits aged 6 weeks were used for culture of BMSCs.METHODS: pDC316-SOX-9 plasmid was used for transfection of rabbit BMSCs by liposome mediated method,Cell transfection included a SOX-9 plasmid transfection group,a blank plasmid group and a blank control group (only treatment of liposome). MAIN OUTCOME MEASURES: Cell morphology; growth activity; The SOX-9 protein expression in rabbit BMSCs were detected by immunohistochemistry,hematoxylin-eosin staining,reserve transcriptase-polymcrase chain reaction (RT-PCR) and enzyme-labeled immunosorbent assay (ELISA). RESULTS: Some cell colonies were detected at 4 days after pDC316-SOX-9 plasmid transfection.Spindle-shaped cells were collected after clone amplification.Cells in the blank control group gradually died over time.Cell activities in the SOX-9 plasmid transfeetion group and the blank plasmid group significantly prolonged,reached a peak at 2 weeks of transfection,and then gradually decreased.At 6 days,BMSCs were yellow in the SOX-9 plasmid transfection group following immunohistochemistry,expressing SOX-9 protein.Hematoxyliu-eosin staining showed many dikaryocytes,rich cell proliferation,similar to chondroblasts.No SOX-9 protein expression and unproductive cell proliferation in the blank plasmid group.SOX-9 mRNA was detected in the SOX-9 plasmid transfection group,but SOX-9 mRNA was not detected in the blank plasmid group and blank control group.SOX-9 levels were significantly higher in the SOX-9 plasmid transfection group than in the blank plasmid group and blank control group at 24,48 and 72 hours,1,2 weeks (P＜ 0.01).CONCLUSION: Rabbit BMSCs were successfully transfected with pDC316-SOX-9 plasmid to enhance cell growth activity and to persistently stably secrete SOX-9 protein,resulting in the differentiation of BMSCs into cartilages.